7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid as an anti-inflammatory compound

ABSTRACT

A 7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid compound, its synthesis, and its use as an anti-inflammatory agent.

BACKGROUND 1. Field

The present disclosure relates to the compound7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid, its synthesis, and its use as an anti-inflammatory agent.

2. Description of the Related Art

The investigation of organic compounds for drug discovery has been arapidly emerging theme in medicinal chemistry. For example, due to theenhanced global encumbrance of cancer in the last decades, extensiveresearch has been devoted to discovering novel selective potentcandidates with the promising ability to either destroy cancer cells orlimit their proliferation.

Similarly, bacterial infection remains a significant threat to humanlife due to its increasing resistance to conventional antibiotics, whichis a growing public health concern. As a result, there is a criticalneed to create new antimicrobial agents with potent anti-drug-resistantmicroorganism activity, as well as associated anti-inflammatoryactivities.

The chemistry of heterocycles lies at the heart of drug discovery. Forexample, arylidene-2-phenyl-1H-imidazol-5(4H)-ones have exhibiteddifferent bioactivities such as immunomodulatory, anti-inflammatory,leishmanicidal, anticancer, and antimicrobials activities. Therefore,the synthesis and bioassay of these types of compounds have absorbedgreat and persistent attention during half of the century.

Furthermore, ciprofloxacin is an antibiotic agent and its derivativeshave exhibited diverse biological activities, such as antibacterial,anti-tuberculosis, antifungal, anti-HIV, anti-malarial, anti-tumor,anti-ischemic, and anti-oxidation activities, as well as inhibitoryurease activity. Various imaging profiles have been developed in thelast years, and the antibacterial properties of ciprofloxacinderivatives have been studied accordingly.

Further, multi-component reactions (MCRs) are economically andenvironmentally beneficial to industry and have attracted muchattention.

Thus, new molecules having desired therapeutic activities and solvingthe aforementioned problems are desired.

SUMMARY

The present subject matter relates to the synthesis of various “drublike” substances having biological activity, as well as such synthesizedcompounds themselves. Hence, herein the present subject matter relatesto synthesis of7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4dihydroquinoline-3-carboxylicacid (4) via a one-pot three-component reaction of4-benzylidene-2-phenyl-1H-imidazol-5(4H)-one 1, formaldehyde 2, andciprofloxacin 3 in ethanol. This synthesized product is likely to havehigh anti-inflammatory activity.

In an embodiment, the present subject matter relates to a7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylic acidcompound having the formula I:

In another embodiment, the present subject matter relates to apharmacuetically acceptable composition comprising a therapeuticallyeffective amount of the7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound and a pharmaceutically acceptable carrier.

In an additional embodiment, the present subject matter relates to amethod of treating inflammation in a patient comprising administering toa patient in need thereof a therapeutically effective amount of the7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound.

In one more embodiment, the present subject matter relates to a methodof making the7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound, the method comprising: stirring a solution of4-benzylidene-2-phenyl-1H-imidazol-5(4H)-one and formaldehyde in ethanolto obtain a reaction mixture; adding ciprofloxacin HCl and triethylamineto the reaction mixture and stirring with reflux; cooling the reactionmixture at room temperature to obtain a solid; and obtaining the7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound.

These and other features of the present subject matter will becomereadily apparent upon further review of the following specification.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a ¹H NMR analysis of the7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

The following definitions are provided for the purpose of understandingthe present subject matter and for construing the appended patentclaims.

Definitions

Throughout the application, where compositions are described as having,including, or comprising specific components, or where processes aredescribed as having, including, or comprising specific process steps, itis contemplated that compositions of the present teachings can alsoconsist essentially of, or consist of, the recited components, and thatthe processes of the present teachings can also consist essentially of,or consist of, the recited process steps.

It is noted that, as used in this specification and the appended claims,the singular forms “a”, “an”, and “the” include plural references unlessthe context clearly dictates otherwise.

In the application, where an element or component is said to be includedin and/or selected from a list of recited elements or components, itshould be understood that the element or component can be any one of therecited elements or components, or the element or component can beselected from a group consisting of two or more of the recited elementsor components. Further, it should be understood that elements and/orfeatures of a composition or a method described herein can be combinedin a variety of ways without departing from the spirit and scope of thepresent teachings, whether explicit or implicit herein.

The use of the terms “include,” “includes”, “including,” “have,” “has,”or “having” should be generally understood as open-ended andnon-limiting unless specifically stated otherwise.

The use of the singular herein includes the plural (and vice versa)unless specifically stated otherwise. In addition, where the use of theterm “about” is before a quantitative value, the present teachings alsoinclude the specific quantitative value itself, unless specificallystated otherwise. As used herein, the term “about” refers to a ±10%variation from the nominal value unless otherwise indicated or inferred.

The term “optional” or “optionally” means that the subsequentlydescribed event or circumstance may or may not occur, and that thedescription includes instances where said event or circumstance occursand instances in which it does not.

It will be understood by those skilled in the art with respect to anychemical group containing one or more substituents that such groups arenot intended to introduce any substitution or substitution patterns thatare sterically impractical and/or physically non-feasible.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood to one of ordinary skill inthe art to which the presently described subject matter pertains.

Where a range of values is provided, for example, concentration ranges,percentage ranges, or ratio ranges, it is understood that eachintervening value, to the tenth of the unit of the lower limit, unlessthe context clearly dictates otherwise, between the upper and lowerlimit of that range and any other stated or intervening value in thatstated range, is encompassed within the described subject matter. Theupper and lower limits of these smaller ranges may independently beincluded in the smaller ranges, and such embodiments are alsoencompassed within the described subject matter, subject to anyspecifically excluded limit in the stated range. Where the stated rangeincludes one or both of the limits, ranges excluding either or both ofthose included limits are also included in the described subject matter.

Throughout the application, descriptions of various embodiments use“comprising” language. However, it will be understood by one of skill inthe art, that in some specific instances, an embodiment canalternatively be described using the language “consisting essentiallyof” or “consisting of”.

“Subject” as used herein refers to any animal classified as a mammal,including humans, domestic and farm animals, and zoo, sports, and petcompanion animals such as household pets and other domesticated animalssuch as, but not limited to, cattle, sheep, ferrets, swine, horses,poultry, rabbits, goats, dogs, cats and the like.

“Patient” as used herein refers to a subject in need of treatment of acondition, disorder, or disease, such as inflammation.

For purposes of better understanding the present teachings and in no waylimiting the scope of the teachings, unless otherwise indicated, allnumbers expressing quantities, percentages or proportions, and othernumerical values used in the specification and claims, are to beunderstood as being modified in all instances by the term “about”.Accordingly, unless indicated to the contrary, the numerical parametersset forth in the following specification and attached claims areapproximations that may vary depending upon the desired propertiessought to be obtained. At the very least, each numerical parametershould at least be construed in light of the number of reportedsignificant digits and by applying ordinary rounding techniques.

The present subject matter relates to the synthesis of various “druglike” substances having biological activity, as well as such synthesizedcompounds themselves. Hence, herein the present subject matter relatesto synthesis of7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4oxo-1,4-dihydroquinoline-3-carboxylicacid (4) via a one-pot three-component reaction of4-benzylidene-2-phenyl-1H-imidazol-5(4H)-one 1, formaldehyde 2, andciprofloxacin 3 in ethanol. This synthesized product is likely to havehigh anti-inflammatory activity.

In an embodiment, the present subject matter relates to a7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound having the formula I:

In certain embodiments, the7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound can be obtained as a yellow solid. In further embodiments,the7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound can have a melting point of about 232° C. to about 234° C.

In another embodiment, the present subject matter relates to apharmaceutically acceptable composition comprising a therapeuticallyeffective amount of the7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound and a pharmaceutically acceptable carrier.

In this regard, the present subject matter is further directed topharmaceutical compositions comprising a therapeutically effectiveamount of the compound as described herein together with one or morepharmaceutically acceptable carriers, excipients, or vehicles. In someembodiments, the present compositions can be used for combinationtherapy, where other therapeutic and/or prophylactic ingredients can beincluded therein.

The present subject matter further relates to a pharmaceuticalcomposition, which comprises a present compound together with at leastone pharmaceutically acceptable auxiliary.

Non-limiting examples of suitable excipients, carriers, or vehiclesuseful herein include liquids such as water, saline, glycerol,polyethylene glycol, hyaluronic acid, ethanol, and the like. Suitableexcipients for nonliquid formulations are also known to those of skillin the art. A thorough discussion of pharmaceutically acceptableexcipients and salts useful herein is available in Remington'sPharmaceutical Sciences, 18th Edition. Easton, Pa., Mack PublishingCompany, 1990, the entire contents of which are incorporated byreference herein.

The present compound is typically administered at a therapeutically orpharmaceutically effective dosage, e.g., a dosage sufficient to providetreatment for cancer or inflammation. Administration of the compound orpharmaceutical compositions thereof can be by any method that deliversthe compound systemically and/or locally. These methods include oralroutes, parenteral routes, intraduodenal routes, and the like.

While human dosage levels have yet to be optimized for the presentcompound, generally, a daily dose is from about 0.01 to 10.0 mg/kg ofbody weight, for example about 0.1 to 5.0 mg/kg of body weight. Theprecise effective amount will vary from subject to subject and willdepend upon the species, age, the subject's size and health, the natureand extent of the condition being treated, recommendations of thetreating physician, and the therapeutics or combination of therapeuticsselected for administration. The subject may be administered as manydoses as is required to reduce and/or alleviate the signs, symptoms, orcauses of the disease or disorder in question, or bring about any otherdesired alteration of a biological system.

In employing the present compound for treatment of inflammation, anypharmaceutically acceptable mode of administration can be used withother pharmaceutically acceptable excipients, including solid,semi-solid, liquid or aerosol dosage forms, such as, for example,tablets, capsules, powders, liquids, suspensions, suppositories,aerosols or the like. The present compounds can also be administered insustained or controlled release dosage forms, including depotinjections, osmotic pumps, pills, transdermal (includingelectrotransport) patches, and the like, for the prolongedadministration of the compound at a predetermined rate, preferably inunit dosage forms suitable for single administration of precise dosages.

The present compounds may also be administered as compositions preparedas foods for humans or animals, including medical foods, functionalfood, special nutrition foods and dietary supplements. A “medical food”is a product prescribed by a physician that is intended for the specificdietary management of a disorder or health condition for whichdistinctive nutritional requirements exist and may include formulationsfed through a feeding tube (referred to as enteral administration orgavage administration).

A “dietary supplement” shall mean a product that is intended tosupplement the human diet and may be provided in the form of a pill,capsule, tablet, or like formulation. By way of non-limiting example, adietary supplement may include one or more of the following dietaryingredients: vitamins, minerals, herbs, botanicals, amino acids, anddietary substances intended to supplement the diet by increasing totaldietary intake, or a concentrate, metabolite, constituent, extract, orcombinations of these ingredients, not intended as a conventional foodor as the sole item of a meal or diet. Dietary supplements may also beincorporated into foodstuffs, such as functional foods designed topromote control of glucose levels. A “functional food” is an ordinaryfood that has one or more components or ingredients incorporated into itto give a specific medical or physiological benefit, other than a purelynutritional effect. “Special nutrition food” means ingredients designedfor a particular diet related to conditions or to support treatment ofnutritional deficiencies.

Generally, depending on the intended mode of administration, thepharmaceutically acceptable composition will contain about 0.1% to 90%,for example about 0.5% to 50%, by weight of the present compound, theremainder being suitable pharmaceutical excipients, carriers, etc.

One manner of administration for the conditions detailed above is oral,using a convenient daily dosage regimen which can be adjusted accordingto the degree of affliction. For such oral administration, apharmaceutically acceptable, non-toxic composition is formed by theincorporation of any of the normally employed excipients, such as, forexample, mannitol, lactose, starch, magnesium stearate, sodiumsaccharine, talcum, cellulose, sodium croscarmellose, glucose, gelatin,sucrose, magnesium carbonate, and the like. Such compositions take theform of solutions, suspensions, tablets, dispersible tablets, pills,capsules, powders, sustained release formulations and the like.

The present compositions may take the form of a pill or tablet and thusthe composition may contain, along with the active ingredient, a diluentsuch as lactose, sucrose, dicalcium phosphate, or the like; a lubricantsuch as magnesium stearate or the like; and a binder such as starch, gumacacia, polyvinyl pyrrolidine, gelatin, cellulose, and derivativesthereof, and the like.

Liquid pharmaceutically administrable compositions can, for example, beprepared by dissolving, dispersing, etc. an active compound as definedabove and optional pharmaceutical adjuvants in a carrier, such as, forexample, water, saline, aqueous dextrose, glycerol, glycols, ethanol,and the like, to thereby form a solution or suspension. If desired, thepharmaceutical composition to be administered may also contain minoramounts of nontoxic auxiliary substances such as wetting agents,emulsifying agents, or solubilizing agents, pH buffering agents and thelike, for example, sodium acetate, sodium citrate, cyclodextrinderivatives, sorbitan monolaurate, triethanolamine acetate,triethanolamine oleate, etc.

For oral administration, a pharmaceutically acceptable non-toxiccomposition may be formed by the incorporation of any normally employedexcipients, such as, for example, pharmaceutical grades of mannitol,lactose, starch, magnesium stearate, talcum, cellulose derivatives,sodium croscarmellose, glucose, sucrose, magnesium carbonate, sodiumsaccharin, talcum, and the like. Such compositions take the form ofsolutions, suspensions, tablets, capsules, powders, sustained releaseformulations and the like.

For a solid dosage form, a solution or suspension in, for example,propylene carbonate, vegetable oils or triglycerides, may beencapsulated in a gelatin capsule. Such diester solutions, and thepreparation and encapsulation thereof, are disclosed in U.S. Pat. Nos.4,328,245; 4,409,239; and 4,410,545, the contents of each of which areincorporated herein by reference. For a liquid dosage form, thesolution, e.g., in a polyethylene glycol, may be diluted with asufficient quantity of a pharmaceutically acceptable liquid carrier,e.g., water, to be easily measured for administration.

Alternatively, liquid or semi-solid oral formulations may be prepared bydissolving or dispersing the active compound or salt in vegetable oils,glycols, triglycerides, propylene glycol esters (e.g., propylenecarbonate) and the like, and encapsulating these solutions orsuspensions in hard or soft gelatin capsule shells.

Other useful formulations include those set forth in U.S. Pat. Nos. Re.28,819 and 4,358,603, the contents of each of which are herebyincorporated by reference.

Another manner of administration is parenteral administration, generallycharacterized by injection, either subcutaneously, intramuscularly, orintravenously. Injectables can be prepared in conventional forms, eitheras liquid solutions or suspensions, solid forms suitable for solution orsuspension in liquid prior to injection, or as emulsions. Suitableexcipients are, for example, water, saline, dextrose, glycerol, ethanol,or the like. In addition, if desired, the pharmaceutical compositions tobe administered may also contain minor amounts of non-toxic auxiliarysubstances such as wetting or emulsifying agents, pH buffering agents,solubility enhancers, and the like, such as for example, sodium acetate,sorbitan monolaurate, triethanolamine oleate, cyclodextrins, etc.

Another approach for parenteral administration employs the implantationof a slow-release or sustained-release system, such that a constantlevel of dosage is maintained. The percentage of active compoundcontained in such parenteral compositions is highly dependent on thespecific nature thereof, as well as the activity of the compound and theneeds of the subject. However, percentages of active ingredient of 0.01%to 10% in solution are employable and will be higher if the compositionis a solid which will be subsequently diluted to the above percentages.The composition may comprise 0.2% to 2% of the active agent in solution.

Nasal solutions of the active compound alone or in combination withother pharmaceutically acceptable excipients can also be administered.

Formulations of the active compound or a salt may also be administeredto the respiratory tract as an aerosol or solution for a nebulizer, oras a microfine powder for insufflation, alone or in combination with aninert carrier such as lactose. In such a case, the particles of theformulation have diameters of less than 50 microns, for example lessthan 10 microns.

In an additional embodiment, the present subject matter relates to amethod of treating inflammation in a patient comprising administering toa patient in need thereof a therapeutically effective amount of the7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound.

In one more embodiment, the present subject matter relates to a methodof making the7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound, the method comprising: stirring a solution of4-benzylidene-2-phenyl-1H-imidazol-5(4H)-one and formaldehyde in ethanolto obtain a reaction mixture; adding ciprofloxacin HCl and triethylamineto the reaction mixture and stirring with reflux; cooling the reactionmixture at room temperature to obtain a solid; and obtaining the7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound.

The present production methods can be further seen by referring to thefollowing Scheme 1:

In an embodiment of the present production methods, the reaction mixturecan be stirred at room temperature for at least about 40 minutes beforethe adding of the ciprofloxacin HCl and triethylandrie.

In another embodiment of the present production methods, after theadding of the ciprofloxacin HCl and triethylannne, the reaction mixturecan be stirred for at least about 1 hour2.

In a further embodiment of the present production methods, the solid canbe collected by filtration, washed with water, dried, and recrystallizedfrom ethanol to obtain the7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound.

In an embodiment of the present production methods, the4-benzylidene-2-phenyl-1H-imidazol-5(4H)-one and formaldehyde can beadded in an about 3:7 molar ratio. Similarly, the ciprofloxacin HCl andtriethylamine can be added in an about 3:4 molar ratio. Likewise, the4-benzylidene-2-phenyl-1H-imidazol-5(4H)-one, formaldehyde,ciprofloxacin HCl, and triethylamine can be added in an about 3:7:3:4molar ratio.

In an additional embodiment of the present production methods, the7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound can be obtained in an about 74% yield.

The following examples relate to various methods of manufacturing thespecific compounds and application of the same, as described herein. Allcompound numbers expressed herein are with reference to the syntheticpathway figures shown above.

EXAMPLES Example 1 Preparation of7-(4((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid

A solution of 4-benzylidene-2-phenyl-1H-imidazol-5(4H)-one (7.44 g,0.003 mol) and formaldehyde (0.2 g, 0.007 mol) was stirred in 50 mlethanol for 40 min, then ciprofloxacin HCl (1.28 g, 0.003 mol), andtriethylarnine (0.100 g, 0.004 mol) were added and the reaction mixturewas stirred with reflux for 1 h. The reaction mixture was allowed tocool at room temperature, the separated solid was filtered off, washedwith water, and recrystallized from ethanol to give the desired product7-(4-((4-benzylidene-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid (4).

Characterization of the prepared compound using ¹ H NMR analysis isshown in FIG. 1 . The elemental analysis can be seen as follows.

7-(4((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid (4):

Pale Yellow solid; Mp. 232-234° C. Yield=74%. FTIR (KBr, cm⁻¹) 3357(OH), 3039 (CHarom.), 2961, 2872 (CHaliph.), 1726 and 1715 (C═O). ¹H-NMR (DMSO-d6/D2O, 400 MHz): 15.12(br. OH, COOH), 8.66(s, 1H, CHarom),7.89(d, J=12.3, 1H, CHarom), 7.59(d, J=7.6, 1H, CHarom), 7.50(m, 3H,CHarom.), 7.45-7.33(m, 8H, CHarom.), 5.30(s, 2H, N—CH₂), 3.82(br. s.,1H, CH cyclopropyl), 3.41(s, 4H, 2 N—CH₂), 3.09(m, 4H, 2 N—CH₂), 1.31(d,J=6.3, 2H, CH₂cyclopropyl), 1.18(br. s, 2H, CH₂cyclopropyl); Analysis:calculated for C₃₄H₃₀FN₅O₄ (670.53): C, 69.02; H, 5.11; N, 11.84%.Found: C, 69.12; H, 5.01; N, 11.74%.

Example 2 Anti-Inflammatory Activity

Bovine Serum Albumin Assay (BSA)

In vitro-inflammatory activities of the tested samples were determinedusing a modified method of the BSA assay reported by Williams et al. BSAsolution (0.4%, w/v) was prepared in Tris Buffered Saline. The pH wasadjusted to 6.4 with glacial acetic acid. Different dilutions of 20, 40,60, 80, and 100 pg/mL of the stock solutions of7-(4((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid (4) were added to test tubes containing 1 mL of 0.4%, w/v BSAbuffer solution. Both negative (solvent) and positive (aspirin) controlswere assayed in a similar manner. The solutions were then heated in awater bath at 72° C. for 10 minutes and cooled for 20 minutes underlaboratory conditions. The turbidity of the solutions (level of proteinprecipitation) was measured at 660 nm in Spectrophotometer (V-770UV-Visible/NIR Spectrophotometer).

The percentage inhibition of precipitation (protein denaturation) wasdetermined relative to the negative control using the followingequation:

${\%{Anti}}‐{{{Denaturation}{Activity}} = \frac{{{Absorbance}{of}{control}} - {{Absorbance}{of}{sample} \times 100}}{{Absorbance}{of}{control}}}$%Anti‐DenaturationActivity = %InhibitionofProteinDenaturation = %Anti‐inflammatoryActivity

The in vitro anti-inflammatory activity study of7-(4((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid (4) showed that the synthesized compound7-(4((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid demonstrated a considerable anti-inflammatory effect. Whereasstandard aspirin at 100 pg/ml showed 90.75% inhibition, the present7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound exhibited 84.78% inhibition. Hence it can potentially beused effectively in the management of inflammation.

It is to be understood that the7-(4((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound, compositions containing the same, and methods of usingand producing the same are not limited to the specific embodimentsdescribed above, but encompasses any and all embodiments within thescope of the generic language of the following claims enabled by theembodiments described herein, or otherwise shown in the drawings ordescribed above in terms sufficient to enable one of ordinary skill inthe art to make and use the claimed subject matter.

We claim:
 1. A7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound having the formula I:


2. The7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound of claim 1, wherein the compound is obtained as a yellowsolid.
 3. The7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound of claim 1, having a melting point of about 232° C. toabout 234° C.
 4. A pharmaceutically acceptable composition comprising atherapeutically effective amount of the7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound of claim 1 and a pharmaceutically acceptable carrier.
 5. Amethod of treating inflammation in a patient comprising administering toa patient in need thereof a therapeutically effective amount of the7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound of claim
 1. 6. A method of making the7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound of claim 1, the method comprising: stirring a solution of4-benzylidene-2-phenyl-1H-imidazol-5(4H)-one and formaldehyde in ethanolto obtain a reaction mixture; adding ciprofloxacin HCl and triethylamineto the reaction mixture and stirring with reflux; cooling the reactionmixture at room temperature to obtain a solid; and obtaining the7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound.
 7. The method of making the7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound of claim 6, wherein the reaction mixture is stirred atroom temperature for at least about 40 minutes before the adding of theciprofloxacin HCl and triedviamine.
 8. The method of making the17-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound of claim 6, wherein after the adding of the ciprofloxacinHCl and triethylamine, the reaction mixture is stirred for at leastabout 1 hour.
 9. The method of making the7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound of claim 6, wherein the solid is collected by filtration,washed with water, dried, and recrystallized from ethanol to obtain the7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound.
 10. The method of making the7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound of claim 6, wherein the4-benzylidene-2-phenyl-1H-imidazol-and formaldehyde are added in anabout 3:7 molar ratio.
 11. The method of making the7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound of claim 6, wherein the ciprofloxacin HCl andtriethylamine are added in an about 3:4 molar ratio.
 12. The method ofmaking the7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound of claim 6, wherein the4-benzylidene-2-phenyl-1H-imidazol-5(4H)-one, formaldehyde,ciprofloxacin HCl, and triethylamine are added in an about 3:7:3:4 molarratio.
 13. The method of making the7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound of claim 9, wherein the7-(4-((4-benzylidene-5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl)methyl)piperazin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylicacid compound is obtained in an about 74% yield.